Fig. 4: Pore-forming gels loaded with GM-CSF, Dox-iRGD, and CpG improve tumor-specific CTL responses and antitumor efficacy against 4T1 triple-negative breast cancer.

a, b 4T1 cells were injected subcutaneously on day 0. Mice were untreated or treated with gels containing Dox-iRGD (100 μg) and CpG (50 μg) or Dox-iRGD (100 μg) alone or CpG (50 μg) alone on day 5. GM-CSF was incorporated in all groups. a Average 4T1 tumor volume of each group over the course of the efficacy study. n = 8 biologically independent animals per group. Statistical analysis was performed using ANOVA with Fisher’s LSD post hoc test. b Kaplan–Meier plots for all groups. n = 5 biologically independent animals per group. Statistical analysis was performed using the log-rank (Mantel–Cox) test. c–g Following 4T1 tumor inoculation on day 0, gels containing Dox-iRGD (200 μg) and CpG (100 μg) or Dox-iRGD (200 μg) alone or CpG (100 μg) alone were injected peritumorally on day 5. c Representative FACS plots of cells isolated from tumor-draining lymph nodes at 4 days post gel injection and restimulated with 4T1 cells. APC-conjugated anti-IFN-γ and Pacific Blue-conjugated anti-CD8 were used for staining. Cells without restimulation were used as control. d Percentage of IFN-γ⁺CD8⁺ cells. e Mean APC fluorescence intensity of CD8⁺ T cells. For d, e, n = 5 biologically independent animals per group. Statistical analysis was performed using ANOVA with Fisher’s LSD post hoc test. f Average 4T1 tumor volume of each group over the course of the efficacy study. Statistical analysis was performed using ANOVA with Fisher’s LSD post hoc test. g Kaplan–Meier plots for all groups. Statistical analysis was performed using the log-rank (Mantel–Cox) test. For f, g, n = 7 biologically independent animals per group. All the numerical data are presented as mean ± SD except for a where mean ± SEM is used. Source data are provided as a Source Data file.